Detection of HPV in Japanese and Chinese oral carcinomas by in situ PCR.

Human papillomavirus (HPV) is established as the cause of almost 100% of cervical carcinomas. However, the association of HPV with oral squamous cell carcinomas (SCCs) is less well understood. We examined the prevalence of HPV in oral SCCs in samples of Japanese and Chinese populations. Using in situ polymerase chain reaction (PCR) analysis (MY09 and MY11 consensus primers), HPV was detected in the nucleus of epithelia and tumor cells in oral lesions. Analysis revealed the specific presence of HPV DNA in all cases of SCC in our Japanese (10/10) and Chinese (10/10) population samples. These results suggest that HPV infection could be one of several risk factors contributing to oral SCC in Japanese and Chinese.

A novel immunohistochemical technique for the detection of human oral carcinomas transplanted in nude mice using mouse monoclonal antibodies.

We have developed a new immunostaining technique specifically for the detection of human tumors transplanted into nude mice using mouse monoclonal antibodies (MoAbs) produced in our laboratory. The formation of a molecular complex consisting of three components (mouse MoAb or hybridoma supernatant, biotin-labeled anti-mouse immunoglobulins, and normal nude mouse serum) markedly reduced background staining and enhanced specific reaction with the transplanted tumors in nude mice. When MoAb production by electrofusion was screened with this new method, the incidence of hybridoma supernatant reactive with sections of transplanted tumor was 2.3 per 100 wells immunostained. These results suggest that production of MoAbs using transplanted tumors is immunohistochemically possible and that this method may provide a new means for developing useful tumor markers.

Cytoplasmic crystalloids in the periodontal tissue of beige mice.

The periodontal tissue of beige mice has been examined by electron microscopy. Specimens of the periodontal tissue from the right mandibular first molars of 10 male beige (bg/bg) and 5 male non-beige (bg/+, controls) mice, were fixed in a mixture of glutaraldehyde and paraformaldehyde, and then decalcified in EDTA. After fixation in osmium tetroxide, the specimens were dehydrated and embedded in epon resin. Ultrathin sections were then stained with uranyl acetate and lead citrate, and observed by electron microscopy. A larger number of cytoplasmic crystalloids were seen in the lysosomes of the histiocytes of the beige mice than in the non-beige mice. These cytoplasmic crystalloids showed striation structures composed of linear densities that were oriented longitudinally, transversely, and obliquely, with about a 4-nm periodicity. The periodontal tissue of the beige mice contained several cells with numerous cytoplasmic crystalloids, whereas there were few cells with fewer crystalloids in the non-beige mice.

Anti-galactocerebroside antibodies in human cerebrospinal fluids determined by enzyme-linked immunosorbent assay (ELISA).

The standard ELISA technique was improved for the detection of antigalactocerebroside antibody in biological fluid. Mouse monoclonal antigalactocerebroside antibody was used to demonstrate specificity and sensitivity of the technique. After optimization of the assay, the usefulness of this measurement for the evaluation of patients with multiple sclerosis was assessed. The presence of antigalactocerebroside antibodies in the cerebrospinal fluid of 20 patients with multiple sclerosis, 10 with other neurological diseases and 10 normal individuals was determined. All the CSF samples from normal individuals were negative. In patients with multiple sclerosis 14 of the 20 samples had elevated levels of antigalactocerebroside antibody, whereas with other neurological diseases 5 out of 10 were positive. Antigalactocerebroside levels were lower in samples from patients during an acute relapse than in those from more chronic cases. These results indicate that the presence of antigalactocerebroside antibody in cerebrospinal fluid is not specific to MS but may reflect previous damage to myelin.

Serum proteins and secretory component in human carious dentin.

By the use of the peroxidase-labeled antibody method, significant localization of IgG, IgA, albumin and transferrin was demonstrated in the deep lesion of 20 carious teeth, where the secretory component was absent. These serum proteins formed a distinct zone, surrounding the overlying, shallow lesion infected with bacteria.

Studies on monoterpene glucosides and related natural products. XXXI. Gas chromatography and gas chromatography-mass spectrometry of iridoid and secoiridoid glucosides.

A total of 33 iridoid and secoiridoid glucosides were detected by gas chromatography on several columns such as OV-1 or OV-17. Representative glucosides were then subjected to gas chromatography-mass spectrometry, giving some characteristic peaks that permitted the discrimination of both types of glucosides from other compounds in most instances. The successful detection of both types of glucosides in several plant extracts showed the applicability of this combination of methods to small amounts of plant materials.